油松菌根根际放线菌的分离与鉴定

为了探讨油松菌根根际放线菌与菌根真菌及油松猝倒病的关系,该实验从油松菌根根际土中分离、纯化得到170株放线菌,采用平板对峙培养法对油松猝倒病病原真菌立枯丝核菌(Rhizoctonia solani)和茄腐皮镰孢霉(Fusarium solani)进行生防放线菌的初筛,得到7株对病原真菌抑制效果较好的放线菌.用初筛选的7株放线菌菌株分别与外生菌根真菌灰鹅膏菌(Amanita vatinata)、灰环粘盖牛肝菌(Suillus laricinus)和血红铆钉菇(Gomphidius viscidus)进行纯培养互作复筛,得到2株放线菌(菌株19#、菌株110#)对外生菌根真菌灰鹅膏菌和血红铆钉菇均具有促生作用.将菌根真菌灰鹅膏菌、放线菌菌株19#、病原真菌立枯丝核菌进行纯培养互作试验表明,菌株19#能协同灰鹅膏菌拮抗立枯丝核菌.经形态特征、培养特征、生理生化特性测定和16SrDNA序列分析,确定菌株19#为苯胺紫链霉菌(Streptomyces mauvecolor).     

松树某些外生菌根真菌对防治油松幼苗猝倒病的作用

从油松根部分离到的外生菌根真菌:牛肝菌之一种(Boletus sp.)和厚环粘盖牛肝菌(Suillus grevillei)除对油松(Pinus tabulaeformis)幼苗猝倒病有防治作用外,还能促进幼苗生长,其显著性水平P=0.01。引起油松幼苗猝倒病的病原菌主要是立枯丝核菌(Rhizoctonia solant)和尖孢镰刀菌(Fusarium oxysporum)。在盆栽接种试验中,用牛肝菌和厚环粘盖牛肝菌接种,油松幼苗发病率比对照降低21.0—36％;在田间接种试验中,松苗发病率比对照降低16.3—25.8％。用扫描电镜对接了种的油松幼苗进行观察,发现在油松营养根表面形成密集的菌丝套,而根尖则没有。用石腊切片观察到围绕在菌根皮层细胞周围的哈蒂氏网。采用500毫升的广口瓶,盛入草炭150克、细砂50克、玉米粉9克、红糖1克、麦芽汁(1.03波美)300毫升的配方,接种牛肝菌菌丝体,置30℃,10天左右产生山蘑菇子实体。     

马尾松与粘盖乳牛肝菌菌根共生体形成过程

对菌根共生机制的研究是对其进行应用的前提,到目前为止,绝大多数外生菌根（ectomycorrhiza,ECM）的建立过程尚不明晰,在一定程度上限制了这些ECM真菌在林业中的应用。本研究以我国南方地区主栽树种之一——马尾松Pinus massoniana和其林下优势ECM真菌——粘盖乳牛肝菌Suillus bovinus为材料,在无菌条件下研究两者菌根共生体形成过程的形态学特征。结果表明马尾松与粘盖乳牛肝菌的共生过程分为2个阶段：（1）预共生阶段,即物理接触之前,粘盖乳牛肝菌可通过释放挥发物和分泌物促进马尾松根系伸长和分枝;（2）共生阶段,又可分为3个时期。接种后第4天,粘盖乳牛肝菌菌丝体开始与马尾松根系接触并形成附着胞进入接触期;第7天菌丝开始侵入根系内部,侵入期开始;第28天菌套和哈氏网形成,即菌套和哈氏网形成期,该时期菌根化根尖开始膨大,随后继续发育至二叉分枝状菌根形成。在发育顺序方面,哈氏网与菌套同步发育,但哈氏网成形早于菌套。以上结果可对后续ECM共生机制的深入研究及马尾松高效菌根化育苗技术的开发提供参考。     

油松菌根促生细菌——荧光假单胞菌的分离与鉴定

为了探讨菌根促生细菌荧光假单胞菌(Pseudomonas fluorescens)与菌根真菌的互作关系,本实验从油松菌根上分离得到36株在紫外灯下产荧光的细菌菌株,以荧光假单胞菌9702作为标准菌株,对分离菌株进行显微观察、生物学鉴定和16S rDNA序列分析,结果确定HDY-8、HDY-9、HDY-20、HDY-35共 4株菌株为荧光假单胞菌,并分别命名为P.fluorescens HDY-8、P.fluorescens HDY-9、P.fluorescens HDY-20、P.fluorescens HDY-35.用这4株细菌菌株分别与外生菌根真菌(ECMF)粘盖牛肝菌(Suillus bovinus)、褐环粘盖牛肝菌(Suillus luteus)和褐黄牛肝菌(Boletus luridus)进行纯培养互作研究.结果表明,只有P.fluorescens HDY-20对3种外生菌根真菌有不同程度的促生作用,并对S.luteus促进效果最好,S.bovinus次之,B.luridus最差;P.fluorescens HDY-20促进S.bovinus、S.luteus和B.luridus菌丝生长的最佳浓度分别为2.4×109 cfu/mL、0.8×109～2.4×109 cfu/mL和0.8×109 cfu/mL,与对照相比S.bovinus和S.luteus的生物量达到极显著差异(P<0.01),B.luridus的达到显著差异(P<0.05),且分别比对照增加6.5%、9.1%和4.3%.     

接种乳黄粘盖牛肝菌和荧光假单胞菌对油松苗生长及猝倒病的影响

在盆栽条件下研究了单接种外生菌根真菌乳黄粘盖牛肝菌(SL)、单接种荧光假单胞菌(PF)、双接种(SL+PF)和不接种菌剂(CK)4个处理对油松苗生长、盆栽土壤指标、油松苗生理生化指标及猝倒病发生的影响.结果显示:(1)不同接种处理的油松苗成活率、生长状况及根系活力均高于对照,且SL+PF促生效果最好,其成活率、苗高、地径、地上干重、总干重及根系活力分别比对照提高22.10%、36.16%、44.86%、124.14%、161.54%和100%.(2)双接种可同时有效提高土壤酶活性和土壤养分含量,土壤脲酶、碱性磷酸酶和蔗糖酶活性均与速效氮、速效磷和有机碳呈显著或极显著正相关.(3)接种病原菌前后油松苗的MDA含量表现为CK>PF>SL>SL+PF,SOD、CAT和PAL活性均为SL+PF>SL>PF>CK,但接种病原菌前后不同处理酶活性增长幅度不同,CAT和PAL活性均以SL+PF增幅最大,分别比对照增长76.18%和112.64%.(4)接种处理的油松苗猝倒病发病率及病情指数均低于对照,且以SL+PF抗病效果最好,分别比对照降低60.94%和70.55%.研究表明,接种菌根真菌可有效提高土壤酶活性和土壤养分含量,增强植株体内的抗氧化酶活性,从而促进油松苗生长,抑制猝倒病的发生,菌根促生菌荧光假单胞菌增强了菌根真菌的作用.     

粘盖牛肝菌不同菌株对Zn2+、Cd2+的吸附及其对油松Zn2+、Cd2+的耐受性

为了探讨同一菌根真菌不同菌株对重金属的耐受性,选用采集于内蒙古阴山山脉不同地区的粘盖牛肝菌(Suillus bovinus)的不同菌株进行研究。首先,在不同浓度Zn^2 、Cd^2 液体培养基中培养菌丝体,以了解各菌株菌丝体对重金属的耐受性及吸附能力,采用烘干法和原予吸收法分别测定菌丝体的生物量和菌丝体、培养液中Zn^2 、Cd^2 含量,结果表明：劈柴沟粘盖牛肝菌在Zn^2 、Cd^2 胁迫条件下,生物量、吸附能力约为其余各菌株的1．5～2倍。其次,为了探明油松(Pinus tabulaeformis)形成菌根后对Zn^2 、Cd^2 胁迫的耐受性及其耐受机理,采用一次性定量浇灌不同浓度Zn^2 、Cd^2 溶液的方法,测定了菌根化油松苗地上、地下生物量及Zn^2 、Cd^2 含量的分配,结果表明：菌根形成后能显著促进油松的生长及对Zn^2 、Cd^2 胁迫的耐受性,并且菌根能够帮助油松吸收基质中大量的Zn^2 、Cd^2 ,根中重金属的含量是茎叶中的2～3倍以上,非菌根苗在重金属浓度稍高(Zn^2 400mg／kg;Cd^2 40mg／kg)时就会死亡。经方差分析及多重比较证实,劈柴沟粘盖牛肝菌对Zn^2 、Cd^2 的耐受性及对油松的促生效果与其它各菌株存在显著的差异,这可能是它通过把吸收的Zn^2 、Cd^2 最大限度地输送到根中的同时,也输送到了茎叶中,使重金属在体内得到一定程度的稀释,使自身免受毒害。     

辽东栎幼苗的外生菌根合成

 在温室花盆中播种辽东栎（Quercus liaotungensis）种子获得辽东栎幼苗,并对幼苗接种外生菌根真菌进行菌根合成试验。所用的外生菌根真菌有：铆钉菇（Gomphidius viscidus）、臭红菇（Russula foetens）、厚环乳牛肝菌（Suillus grevillei）、褐环乳牛肝菌（S. luteus）、彩色豆马勃（Pisolithus tinctorius）、美味牛肝菌（Boletus edulis）、赭丝膜菌（Cortinarius russus）、土生空团菌（Cenococ     

红花尔基樟子松优良抗旱菌树组合的筛选

为筛选红花尔基樟子松（Pinus sylvestris var.mongolica）优良抗旱菌树组合,采用樟子松林下5个优势外生菌根真菌菌株为接种体,分别对5个月龄樟子松实生苗进行人工接种,接种8个月后观察菌根侵染情况及菌根形态,并在非干旱胁迫和干旱胁迫条件下测定不同菌树组合的生长和生理生化指标。结果表明：5个乡土菌种均能成功侵染樟子松并形成典型的外生菌根;干旱胁迫下,菌根化樟子松的各项生长指标均显著高于对照（P<0.05）,且接种粘盖牛肝菌（Suillus bovinus）具有最高的菌根侵染率、苗高、地上及地下生物量和根茎比;外生菌根共生体可通过提高植物SOD活性与POD活性,同时降低MDA含量来提高樟子松对干旱的耐受力;干旱胁迫下,所有接种处理苗木的萎蔫时间较对照处理均推迟,推迟时间最长的是粘盖牛肝菌接种处理,较对照推迟96.3 h;另外,接种处理均能显著延长宿主临界致死时间,尤其是接种粘盖牛肝菌可延长113.8 h。因此,可以认为粘盖牛肝菌与樟子松是一个较为理想的抗旱菌树组合。     

外生菌根真菌对杉木的接种效应

对商南县2年生杉木苗木进行乳牛肝菌(Suillus bovines)、褐环乳牛肝菌(Suillus luteus)和褐黄牛肝菌(Boletus luridus)3种外生菌根真菌的接种试验.结果显示,外生菌根真菌对杉木苗木成活率、苗高和地径生长均有明显的促进作用,能明显提高苗木的菌根化水平,增加苗木体内叶绿素含量,促进其游离脯氨酸含量的累积,降低丙二醛含量.对不同坡向杉木苗木各项生长指标的分析显示,在杉木人工造林中,阳坡的综合效应明显优于阴坡.乳牛肝菌和褐黄牛肝菌可作为一种生物肥料,在杉木人工造林中应用.     

两种乳牛肝菌属菌根食用菌与马尾松的共生特征

菌根食用菌（edible mycorrhizal fungi,EMF）是兼具生态价值和经济价值的大型真菌,其对林木健康、森林生态系统的稳定性及林下经济的发展至关重要。本研究以贵州地区马尾松Pinus massoniana林2种乳牛肝菌属Suillus EMF为研究对象,研究了其子实体及其与马尾松形成的菌根的形态特征,调查了其子实体的发生规律及其与温度和降雨的关系。研究结果表明：粘盖乳牛肝菌S. bovinus和褐环乳牛肝菌S. luteus可同时在马尾松林发生。粘盖乳牛肝菌和褐环乳牛肝菌与马尾松形成的菌根均呈珊瑚状,其中粘盖乳牛肝菌形成的菌根表面光滑无根外菌丝但具有根外菌索,褐环乳牛肝菌形成的菌根具有密集的根外菌丝但无根外菌索。粘盖乳牛肝菌和褐环乳牛肝菌子实体具有较长的发生期、产量较大,且具有错峰发生的特点。相对于降雨,温度是这2种乳牛肝菌子实体发生的主要限制因素。     

两种外生菌根真菌在辽东栎幼苗上的混合接种效应

 辽东栎（Quercus liaotungensis）是中国暖温带落叶阔叶林的主要优势树种之一。铆钉菇（Gomphidius viscidus）和臭红菇（Russula foetens）是在自然环境中与其共生形成外生菌根的真菌。在前期工作证明铆钉菇和臭红菇接种对辽东栎幼苗生长有明显促进作用并以两菌种混合接种效果较好的基础上,探讨了不同的接种量和两个菌种不同比例的混合接种对辽东栎幼苗生长和氮、磷养分的影响。对应于试验的12、18、24 g·pot-1 3个接种量处理,随着接种量的增加,辽东栎幼苗的菌根侵染率增加,铆钉菇菌根的比例增加,而臭红菇菌根的比例减少。辽东栎幼苗的生物量、株高、净光合速率和全株的全氮、全磷含量,均以18 g·pot-1接种量的最高。当接种物中铆钉菇∶臭红菇的比例分别为2∶1、1∶1、1∶2时,辽东栎幼苗的菌根侵染率分别为96.54%、91.02%、92.13%,但彼此间差异不显著。随着接种物中铆钉菇比例的减少,铆钉菇菌根所占比例由42.49%降为23.33%,而臭红菇菌根的比例由57.51%增加为76.67%。辽东栎幼苗的生物量和净光合速率均是以接种比例为1∶1的最高。接种比例为1∶1的辽东栎幼苗的全氮含量也是最高的,并且与另两种接种比例处理的差异显著,而对于全株的全磷含量,则是随着接种物中臭红菇的比例增加而增加。     

接种外生菌根对辽东栎幼苗生长的影响

辽东栎 (Quercusliaotungensis)是中国特有的栎林树种 ,也是中国暖温带落叶阔叶林的主要优势树种之一。铆钉菇 (Gomphidiusviscidus)和臭红菇 (Russulafoetens)是在自然环境中与其共生形成外生菌根的真菌。在温室花盆中播种辽东栎种子获得辽东栎幼苗 ,并对幼苗接种铆钉菇和臭红菇合成外生菌根 ,比较了有菌根和无菌根辽东栎幼苗生长、光合蒸腾特性、氮磷含量的差异。外生菌根对辽东栎幼苗的生长有明显的促进作用 ,有菌根幼苗的生物量、株高、净光合速率和水分利用效率高于无菌根幼苗 ,蒸腾速率则相反。有菌根幼苗的氮磷含量分别为无菌根幼苗的 1.7倍和 2 .2倍 ,外生菌根的合成还改变了氮磷在幼苗器官间的分配比例 ,与无菌根幼苗相比 ,有菌根幼苗茎中的氮磷减少 ,而叶片中的磷显著增加。同时接种铆钉菇和臭红菇的生长促进效果优于单独接种。     

接种外生菌根对辽东栎幼苗生长的影响

 辽东栎（Quercus liaotungensis）是中国特有的栎林树种,也是中国暖温带落叶阔叶林的主要优势树种之一。铆钉菇（Gomphidius viscidus）和臭红菇（Russula foetens）是在自然环境中与其共生形成外生菌根的真菌。在温室花盆中播种辽东栎种子获得辽东栎幼苗,并对幼苗接种铆钉菇和臭红菇合成外生菌根,比较了有菌根和无菌根辽东栎幼苗生长、光合蒸腾特性、氮磷含量的差异。外生菌根对辽东栎幼苗的生长有明显的促进作用,有菌根幼苗的生物量、株高、净光合速率和水分利用效率高于无菌根幼苗,蒸腾速率则相反。有菌根幼苗的氮磷含量分别为无菌根幼苗的1.7倍和2.2倍,外生菌根的合成还改变了氮磷在幼苗器官间的分配比例,与无菌根幼苗相比,有菌根幼苗茎中的氮磷减少,而叶片中的磷显著增加。同时接种铆钉菇和臭红菇的生长促进效果优于单独接种。     

野外模拟酸雨胁迫下接种外生菌根真菌对马尾松幼苗的缓解作用

外生菌根真菌能够提高宿主植物对外界环境胁迫的抵抗力。主要探讨野外条件下外生菌根真菌对酸雨胁迫下马尾松(Pinus massoniana)幼苗生长、养分元素以及表层土壤的影响,以期为酸雨严重区马尾松林恢复提供科学依据。以2年生马尾松幼苗为材料,采用原位试验,共设置6个处理:p H5.6(对照)处理未接种、对照处理接种、p H4.5酸雨处理未接种、p H4.5酸雨处理接种、p H3.5酸雨处理未接种、p H3.5酸雨处理接种。研究表明:(1)酸雨处理与对照处理相比显著降低了非菌根苗总生物量及各部位生物量(根、茎、叶),对株高无显著影响,接种外生菌根真菌可以缓解酸雨对马尾松幼苗生长的不利影响;(2)与对照处理相比,酸雨处理的非菌根苗的针叶中N、P、Ca含量升高,Mg含量降低,根系中N、P、Ca含量降低,Mg含量随p H的降低先升高后降低。接种外生菌根真菌显著提高了p H3.5酸雨处理的马尾松幼苗根系中N、P、Ca、Mg含量,而对针叶中N、P、Ca、Mg含量无显著影响。(3)在非菌根土壤中,p H3.5酸雨处理与对照处理相比显著降低了土壤中有机质、速效磷、速效钾、可溶性碳、可溶性氮、铵态氮、硝态氮含量,而接种外生菌根真菌显著提高了上述指标。酸雨对土壤阳离子交换量无显著影响。总而言之,接种外生菌根真菌促进了酸雨处理的马尾松幼苗生长、缓解了酸雨对马尾松幼苗养分元素和表层土壤的不利影响,由此可见接种外生菌根真菌是减轻酸雨对马尾松危害的一个重要途径。     

Mycorrhizal synthesis of four ectomycorrhizal fungi in potted <Emphasis Type="Italic">Populus maximowiczii</Emphasis> seedlings

Ectomycorrhizal (ECM) syntheses between four ECM fungi, Laccaria amethystina, Hebeloma mesophaeum, Thelephora terrestris, and Tomentella sp., and Populus maximowiczii seedlings that are known to form ECM at a denuded area of Mt. Usu were performed in volcanic debris in a controlled growth chamber. The percentage of ECM colonization and seedling growth were determined 3 months after inoculation. Seedlings were successfully colonized by the inoculated ECM fungi with low contamination ratios. Seedling height and biomass were larger in the inoculated seedlings than in the control, although the effects of inoculation on seedling growth varied with the ECM fungus.     

Induced hydrolytic enzymes of ectomycorrhizal fungi against pathogen Rhizoctonia solani

Interactions between ectomycorrhizal fungi (Suillus laricinus, S. tomentosus, Amanita vaginata and Gomphidius viscidus) and the pathogen Rhizoctonia solani in co-culture were studied using both light and scanning electron microscopy. S. laricinus, S. tomentosus and A. vaginata inhibited the growth of the pathogen. Moreover, A. vaginata exhibited coiling around and penetration of the hyphae into R. solani was observed in the interaction zone. Furthermore, the production of chitinases, beta-1,3-glucanases and beta-glucosidases by these ectomycorrhizal fungi on colloidal chitin or cell walls of R. solani was evaluated: chitinases were not induced by colloidal chitin but all three enzymes were induced by R. solani cell walls. No correlation between inhibition rate and production of lytic enzymes was found.     

Growth response of Afzelia africana Sm. seedlings to ectomycorrhizal inoculation in a nutrient-deficient soil

 The growth and mineral nutrition responses of seedlings of two provenances of Afzelia africana Sm. from Senegal and Burkina Faso, inoculated with four ectomycorrhizal (ECM) fungi (Scleroderma spp. and an unidentified isolate) from the same regions were assessed in a pot experiment in a savanna ECM-propagule-free soil deficient in NPK. There was little variation in the ability of the different fungal species to colonize roots of either provenance of A. africana or to produce external hyphal in soil. Root colonization by ECM fungi and their hyphal development were not related to mineral nutrition or ECM dependency. Differences in P, N, Mg and Ca concentrations in the leaves of inoculated and non-inoculated Afzelia seedlings were not always associated with production of biomass. Only leaf K concentration increased in both provenances after ECM inoculation. However, the Burkina Faso provenance responded better to inoculation with the two fungal isolates than the Senegal provenance in terms of biomass production. This was due to stimulation of root dry weight of the Burkina Faso provenance. Therefore, the hypothesis arises that non-nutritional rather than nutritional effects explain the contribution of ECM inoculation to the growth of A. africana seedlings. Accepted: 27 April 1999     

Effects of VA mycorrhiza on the growth of cacao seedlings under nursery conditions in Venezuela

The effects of vesicular-arbuscular mycorrhizae inoculation on the growth of cacao seedlings (Theobroma cacao var. Ocumare 60) grown for 5 months in a nursery were studied. The effects of introduced VAM fungi (Glomus occultum, Acaulospora appendicula, Glomus manihotis, Acaulospora morrowae and Scutellospora pellucida) in soils treated with copper oxychloride or methyl bromide, were compared with the indigenous VAM and with their respective non-inoculated controls.Cacao seedlings responded well to indigenous VAM fungi, which included Scutellospora calospora as the dominant species, inducing increases significant in height, dry weight and foliar uptake of P, Cu and Zn in relation to the sterile control. G. occultum and A. appendicula increased the height of cacao seedlings but to an extent not yet statistically significant through the duration of the experiment. S. pellucida and A. appendicula doubled the phosphorus uptake of cacao seedlings. The methyl bromide sterilization induced the lowest Cu and Zn uptake in the cacao seedlings but this effect was ameliorated with VAM inoculation. Manifest deficiency symptoms were not observed in the treatments.Copper oxychloride treatment depressed growth to the same level as the sterile control although its residual effects did not kill VAM. It could change the competitive relations among the VAM species and in this case seemed to affect adversely the more efficient native fungi.The influence of the presence and long permanence of large cotyledons in cacao on the results obtained is discussed.     

Ectomycorrhizal pre-inoculation of <Emphasis Type="Italic">Pinus hartwegii</Emphasis> and <Emphasis Type="Italic">Abies religiosa</Emphasis> is replaced by native fungi in a temperate forest of central Mexico

Few studies have focused on analyzing the effect of native inoculated ectomycorrhizal (ECM) fungal strains on seedlings under field conditions in temperate forests. However, it is crucial to verify that the positive effects of ECM under nursery conditions also occur in the field, favoring their performance. We evaluated the short-term effect of ECM on three-year-old seedlings of Pinus hartwegii and Abies religiosa in central Mexico by subjecting them to four treatments: inoculation with Inocybe splendens, inoculation with Suillus brevipes (both native strains), inoculation with forest soil, and non-inoculated plants. Percentage of ECM colonization, plant growth (shoot height and stem diameter), and physiological (osmotic potential, stomatal conductance, CO₂ assimilation and water use efficiency) responses were evaluated. We found that these two ECM species were partial (P. hartwegii) or totally (A. religiosa) replaced after one and a half years in the field. P. hartwegii seedlings increased their water use efficiency during the dry season, but in A. religiosa seedlings, a clear strategy for avoiding water stress was not detected. This ECM replacement had a negative effect on the physiological performance Of A. religiosa. Our results emphasize the importance of selecting compatible fungal-host species combinations for nursery inoculation and of using sources of inoculum adapted to the environmental conditions of the transplant site, ensuring root colonization prior to field transplanting to minimize seedling mortality due to water stress.